Localization and possible role of two different alpha v beta 3 integrin conformations in resting and resorbing osteoclasts

2002 ◽  
Vol 115 (14) ◽  
pp. 2919-2929 ◽  
Author(s):  
Roberta Faccio ◽  
Maria Grano ◽  
Silvia Colucci ◽  
Antonello Villa ◽  
Gianluigi Giannelli ◽  
...  
Keyword(s):  
Bone Resorption ◽  
Membrane Receptors ◽  
Sealing Zone ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Alpha V Beta 3 ◽  
Osteoclast Survival ◽  
Human Osteoclasts

Integrins are membrane receptors that mediate interactions between cells and the extracellular matrix. We recently showed that the osteoclast integrinα vβ3 exists in two different conformations,so-called `basal' and `activated', with each exhibiting a distinct function. In this study we demonstrate that, in non-resorbing osteoclasts, the`activated' form of αvβ3 accumulates in the motile areas of the plasma membrane. During bone resorption this conformation is prevalent in the ruffled membrane, whereas the `basal' form ofα vβ3 is also present in the sealing zone. Moreover, hepatocyte growth factor (HGF) and macrophage colony stimulating factor (M-CSF), two molecules involved in osteoclastogenesis and osteoclast survival, modulate αvβ3 conformation in vitro. Preincubation with HGF or M-CSF induces a shift of conformation ofα vβ3 in primary human osteoclasts (OCs) and in the osteoclast-like cell line (GCT 23). Activated integrin promotes osteoclast migration to the αvβ3 ligand osteopontin and enhances bone resorption. Thus, HGF and M-CSF modulate theα vβ3 conformational states required for osteoclast polarization and resorption. The capacity of growth factors to alter the affinity of αvβ3 toward its ligands offers a potential explanation for the diverse responses of osteoclasts to the same ligand.

scholarly journals Macrophage colony-stimulating factor induces substantial osteoclast generation and bone resorption in human bone marrow cultures

Blood ◽  
1996 ◽  
Vol 88 (7) ◽  
pp. 2531-2540 ◽  
Author(s):  
U Sarma ◽  
AM Flanagan
Keyword(s):  
Bone Resorption ◽  
Vitamin D3 ◽  
Osteoclast Formation ◽  
Dependent Manner ◽  
Colony Stimulating Factor ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Human Osteoclasts ◽  
Stimulating Factor

Macrophage colony-stimulating factor (M-CSF) is essential for murine osteoclast formation and its role in human hematopoiesis in vitro is not fully defined. Therefore, we have investigated the effect of M-CSF on the formation of human osteoclasts in vitro. M-CSF was found to induce substantial bone resorption and osteoclast formation in a dose-responsive and time-dependent manner above that induced by 1,25 dihydroxyvitamin D3 (1,25 vitamin D3) in cultures of human bone marrow (BM) stromal cells sedimented onto devitalized bone. By day 14 there was a mean of approximately 50% of the surfaces of the bone slices resorbed compared with only 6% in cultures treated with 1,25 vitamin D3 alone. Osteoclasts were identified as 23c6+ cells (an antibody that recognizes the vitronectin receptor), 87.5% of which coexpressed the calcitonin receptor. The number of 23c6+ cells correlated strongly with bone resorption spatially, and in a dose-responsive and time-dependent manner; the correlation coefficient in cultures treated with 1,25 vitamin D3 alone was 0.856 and those treated with both M-CSF and 1,25 vitamin D3 was 0.880. Granulocyte-macrophage colony-stimulating factor, IL-1 beta, IL-3, IL-6, tumor necrosis factor-alpha, transforming growth factor-beta, leukemia inhibitory factor, and IL-11 did not increase bone resorption above that in 1,25 vitamin D3-treated cultures. We also found that 1,25 vitamin D3 increased, to a minor but significant degree, both bone resorption and the concentration of M-CSF in the culture supernatants above that in vehicle-treated cultures, indicating that M-CSF is present in our BM cultures, but that there is insufficient to induce substantial osteoclast formation. These results define a critical role for M-CSF in the formation of human osteoclasts.

Role of alpha(v)beta(3) integrin in osteoclast migration and formation of the sealing zone

1999 ◽  
Vol 112 (22) ◽  
pp. 3985-3993 ◽  
Author(s):  
I. Nakamura ◽  
M.F. Pilkington ◽  
P.T. Lakkakorpi ◽  
L. Lipfert ◽  
S.M. Sims ◽  
...  
Keyword(s):  
Bone Resorption ◽  
Cell Attachment ◽  
Leading Edge ◽  
In Vivo Studies ◽  
Sealing Zone ◽  
Osteoclast Function ◽  
Alpha V Beta 3

The alpha(v)beta(3) integrin is abundantly expressed in osteoclasts and has been implicated in the regulation of osteoclast function, especially in cell attachment. However, in vivo studies have shown that echistatin, an RGD-containing disintegrin which binds to alpha(v)beta(3), inhibits bone resorption without changing the number of osteoclasts on the bone surface, suggesting inhibition of osteoclast activity. The objective of this study was to examine how occupancy of alpha(v)beta(3) integrins inhibits osteoclast function, using primary rat osteoclasts and murine pre-fusion osteoclast-like cells formed in a co-culture system. We show that: (1) echistatin inhibits bone resorption in vitro at lower concentrations (IC(50)= 0.1 nM) than those required to detach osteoclasts from bone (IC(50) approximately 1 microM); (2) echistatin (IC(50)= 0.1 nM) inhibits M-CSF-induced migration and cell spreading of osteoclasts; (3) alpha(v)beta(3) integrins are localized in podosomes at the leading edge of migrating osteoclasts, whereas, with echistatin treatment (0.1 nM), alpha(v)beta(3) disperses randomly throughout the adhesion surface; and (4) when bone resorption is fully inhibited with echistatin, there is visible disruption of the sealing zone (IC(50)= 13 nM), and alpha(v)beta(3) visualized with confocal microscopy re-distributes from the basolateral membranes to intracellular vesicular structures. Taken together, these findings suggest that alpha(v)beta(3) integrin plays a role in the regulation of two processes required for effective osteoclastic bone resorption: cell migration (IC(50)= 0.1 nM) and maintenance of the sealing zone (IC(50) approximately 10 nM).

scholarly journals The Alternative Faces of Macrophage Generate Osteoclasts

2016 ◽  
Vol 2016 ◽  
pp. 1-9 ◽  
Author(s):  
N. Lampiasi ◽  
R. Russo ◽  
F. Zito
Keyword(s):  
Macrophage Polarization ◽  
Giant Cells ◽  
Gm Csf ◽  
Receptor Activator ◽  
Inflammatory Stimuli ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Alternatively Activated

The understanding of how osteoclasts are generated and whether they can be altered by inflammatory stimuli is a topic of particular interest for osteoclastogenesis. It is known that the monocyte/macrophage lineage gives rise to osteoclasts (OCs) by the action of macrophage colony stimulating factor (M-CSF) and receptor activator of nuclear factor-kB ligand (RANKL), which induce cell differentiation through their receptors, c-fms and RANK, respectively. The multinucleated giant cells (MGCs) generated by the engagement of RANK/RANKL are typical OCs. Nevertheless, very few studies have addressed the question of which subset of macrophages generates OCs. Indeed, two main subsets of macrophages are postulated, the inflammatory or classically activated type (M1) and the anti-inflammatory or alternatively activated type (M2). It has been proposed that macrophages can be polarizedin vitrotowards a predominantly M1 or M2 phenotype with the addition of granulocyte macrophage- (GM-) CSF or M-CSF, respectively. Various inflammatory stimuli known to induce macrophage polarization, such as LPS or TNF-α, can alter the type of MGC obtained from RANKL-induced differentiation. This review aims to highlight the role of immune-related stimuli and factors in inducing macrophages towards the osteoclastogenesis choice.

Effect of macrophage colony-stimulating factor on in vitro osteoclast generation and bone resorption.

Endocrinology ◽  
1992 ◽  
Vol 130 (1) ◽  
pp. 437-442 ◽  
Author(s):  
V A Corboz ◽  
M G Cecchini ◽  
R Felix ◽  
H Fleisch ◽  
G van der Pluijm ◽  
...  
Keyword(s):  
Bone Resorption ◽  
Colony Stimulating Factor ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor

scholarly journals Macrophage colony-stimulating factor stimulates survival and chemotactic behavior in isolated osteoclasts.

1993 ◽  
Vol 178 (5) ◽  
pp. 1733-1744 ◽  
Author(s):  
K Fuller ◽  
J M Owens ◽  
C J Jagger ◽  
A Wilson ◽  
R Moss ◽  
...  
Keyword(s):  
Bone Resorption ◽  
Osteoclast Formation ◽  
Cell Polarization ◽  
Colony Stimulating Factor ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
And Migration ◽  
Established Role ◽  
Stimulating Factor

Macrophage colony-stimulating factor (M-CSF) is known to play an important role in osteoclast formation. However, its actions on mature cells have not been fully characterized. We now report that M-CSF dramatically stimulates osteoclastic motility and spreading; osteoclasts responded to a gradient of M-CSF with orientation, and random cell polarization occurred after isotropic exposure. M-CSF also supported the survival of osteoclasts by preventing apoptosis. Paradoxically, M-CSF inhibits bone resorption by isolated osteoclasts. We found that this was effected predominantly by reduction in the number of excavations. Thus, M-CSF showed a propensity to suppress resorption through a reduction in the proportion of cells that were resorbing bone. Our data suggest that apart from the established role of M-CSF in the provision of precursors for osteoclastic induction, a major role for M-CSF in bone resorption is to enhance osteoclastic survival, migration, and chemotaxis. It seems appropriate that during these processes resorptive functions should be suppressed. We suggest that M-CSF continues to modulate osteoclastic activity once osteoclasts are on resorptive sites, through regulation of the balance between resorption and migration, such that not only the quantity, but the spatial pattern of resorption can be controlled by adjacent M-CSF-secreting cells of osteoblastic lineage.

scholarly journals A Combination of Osteoclast Differentiation Factor and Macrophage-Colony Stimulating Factor Is Sufficient for both Human and Mouse Osteoclast Formation in Vitro

Endocrinology ◽  
1998 ◽  
Vol 139 (10) ◽  
pp. 4424-4427 ◽  
Author(s):  
Julian M. W. Quinn ◽  
Jan Elliott ◽  
Matthew T. Gillespie ◽  
T. John Martin
Keyword(s):  
Bone Resorption ◽  
Stromal Cells ◽  
Osteoclast Differentiation ◽  
Colony Stimulating Factor ◽  
Human Osteoclast ◽  
Osteoclast Differentiation Factor ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor

Abstract Both human and murine osteoclasts can be derived in vitro from hematopoietic cells or monocytes that are cocultured with osteoblasts or marrow-derived stromal cells. The osteoclastogenic stimulus provided by murine osteoblasts and marrow-derived stromal cells is now known to be mediated by osteoclast differentiation factor (ODF), a membrane-bound tumor necrosis factor-related ligand. This study demonstrates that mouse spleen cells and monocytes form osteoclasts when cultured in the presence of macrophage-colony stimulating factor (M-CSF) and a soluble form of murine ODF (sODF). Numerous multinucleated osteoclasts expressing tartrate resistant acid phosphatase (TRAP) and calcitonin receptor (CTR) formed within 7 days of culture and engaged in extensive lacunar bone resorption. Osteoclast number and bone resorption area was dependent on sODF concentration. Long-term cultured human monocytes also formed bone resorbing osteoclasts in response to co-stimulation by sODF and M-CSF, although this required more than 11 days in culture. This human osteoclast differentiation was strongly inhibited by granulocyte-macrophage colony stimulating factor. This study further characterises murine osteoclast differentiation caused by sODF and M-CSF co-stimulation in vitro, and shows that the same co-stimulation causes human osteoclast differentiation to occur. We propose that this methodology can be employed to investigate the direct effects of cytokines and other factors on human osteoclast differentiation.

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